title: Interaction of the WD40 domain of a myoinositol polyphosphate 5-phosphatase with SnRK1 links inositol, sugar, and stress signaling.
abstract: In plants, myoinositol signaling pathways have been associated with several stress, developmental, and physiological processes, but the regulation of these pathways is largely unknown. In our efforts to better understand myoinositol signaling pathways in plants, we have found that the WD40 repeat region of a myoinositol polyphosphate 5-phosphatase (5PTase13; At1g05630) interacts with the sucrose nonfermenting-1-related kinase (SnRK1.1) in the yeast two-hybrid system and in vitro. Plant SnRK1 proteins (also known as AKIN10/11) have been described as central integrators of sugar, metabolic, stress, and developmental signals. Using mutants defective in 5PTase13, we show that 5PTase13 can act as a regulator of SnRK1 activity and that regulation differs with different nutrient availability. Specifically, we show that under low-nutrient or -sugar conditions, 5PTase13 acts as a positive regulator of SnRK1 activity. In contrast, under severe starvation conditions, 5PTase13 acts as a negative regulator of SnRK1 activity. To delineate the regulatory interaction that occurs between 5PTase13 and SnRK1.1, we used a cell-free degradation assay and found that 5PTase13 is required to reduce the amount of SnRK1.1 targeted for proteasomal destruction under low-nutrient conditions. This regulation most likely involves a 5PTase13-SnRK1.1 interaction within the nucleus, as a 5PTase13:green fluorescent protein was localized to the nucleus. We also show that a loss of function in 5PTase13 leads to nutrient level-dependent reduction of root growth, along with abscisic acid (ABA) and sugar insensitivity. 5ptase13 mutants accumulate less inositol 1,4,5-trisphosphate in response to sugar stress and have alterations in ABA-regulated gene expression, both of which are consistent with the known role of inositol 1,4,5-trisphosphate in ABA-mediated signaling. We propose that by forming a protein complex with SnRK1.1 protein, 5PTase13 plays a regulatory role linking inositol, sugar, and stress signaling.
authors.affiliation: Department of Biochemistry, Virginia Tech, Blacksburg, Virginia 24061.
authors.forename: Elitsa A
authors.initials: EA
authors.lastname: Ananieva
authors.forename: Glenda E
authors.initials: GE
authors.lastname: Gillaspy
authors.forename: Amanda
authors.initials: A
authors.lastname: Ely
authors.forename: Ryan N
authors.initials: RN
authors.lastname: Burnette
authors.forename: F Les
authors.initials: FL
authors.lastname: Erickson
mesh_terms: D000040:Abscisic Acid / Q000494:pharmacology; D020816:Amino Acid Motifs; D017360:Arabidopsis / Q000187:drug effects / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism; D004795:Enzyme Stability / Q000235:genetics; D015870:Gene Expression; D005816:Genetic Complementation Test; D005947:Glucose / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000032:analysis; D007295:Inositol Phosphates / Q000378:metabolism*; D016254:Mutagenesis, Insertional; D010744:Phosphoric Monoester Hydrolases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010802:Phylogeny; D010937:Plant Growth Regulators / Q000494:pharmacology; D018517:Plant Roots / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D025941:Protein Interaction Mapping; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000032:analysis; D015398:Signal Transduction; D013312:Stress, Physiological
chemical_list: D000040:Abscisic Acid; D029681:Arabidopsis Proteins; D005947:Glucose; D049452:Green Fluorescent Proteins; D007295:Inositol Phosphates; D010744:Phosphoric Monoester Hydrolases; D010937:Plant Growth Regulators; D017346:Protein Serine-Threonine Kinases; D011993:Recombinant Fusion Proteins; C052128:inositol 3-phosphate; C531469:At3g01090 protein, Arabidopsis; C499855:5PTase13 protein, Arabidopsis